Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda

Endpoint summary

Description of key information

In a repeated dose toxicity study the by oral route, the test item was administered by drinking water to rats (10 animals/sex/dose) at doses of 0; 100; 500 and 2500 ppm during 5-weeks. Under the test conditions, the test item Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda induced only a slight reduction in body weight gain in males at the highest dose tested (2500 ppm). In the absence of other test item-related changes, this variation was considered of no toxicological relevance. The NOAEL was therefore considered to be 2500 ppm (i.e. 317.05 and 335.22 mg/kg b.w./day in males and females, respectively). No classification as STOT-RE is thus required according to the Regulation (EC) 1272/2008 (CLP) and as harmful after repeated dose exposure according to the Directive 67/548/EEC.
In a repeated dose inhalation toxicity study, groups of Wistar rats (5/sex) were exposed by inhalation route to the test substance as dust aerosol for 6 hours per day on 5 days per week for 4 weeks (28-day test) to head/nose at concentrations of 0, 0.8, 8, 80 mg/m³ (nominal concentrations) or 0, 0.86, 8.1, 82 mg/m³ (analytical concentrations). Based on the results of this study, 82 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) systemic based on the absence of toxicologically significant sytemic effects; 0,86  mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) local and 8.1 mg/m³ of test item was established as the Lowest Observed Adverse Effect Level (LOAEL) local based on effects on the lung weight. Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC. 

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is sufficiently documented. Few deviations from the current guidelines were noted.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

5-week treatment period instead of 4. Histopathology was conducted only on a selection of organs collected at necropsy. No analytical verifications of the doses administered.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:SPF. BOR: WISW (SPF CPB) of the breeder Winkelmann (Borchen)
- Age at study initiation: the rats were 5-6 weeks old
- Weight at study initiation: males: a mean body weight of 105 g, and female a mean body weight of 96 g.
- Fasting period before study: no
- Housing: The animals were placed in a traditional way each cage Makrolon Type II (and Gönnert SPIEGEL, 1961) on wood pellets free of dust.
- Diet (e.g. ad libitum): During the test, the animals received during the weekly cage change their weekly amount of food in the form of granules costs (Altromin 1324 Manufacturer: Altrogge GmbH, Lage).
- Water (e.g. ad libitum): The animals had free access to deionized drinking water, which was diluted test substancel.
- Acclimation period: the animals were acclimated for a period of 12 days before the beginning of the treatment period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 10%
- Air changes (per hr): no
- Photoperiod (hrs dark / hrs light): rhythm day / night 12 hours (artificial light from 6 to 18 pm CET)

IN-LIFE DATES: no data available

Route of administration:

oral: drinking water

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance was diluted once a week in deionized water. To dissolve the deposits present at the bottom, it was heated to approx. 50 ° C. The control subjects received only de-ionized water.

VEHICLE
- de-ionized water
- Concentration in vehicle: 100, 500, 2500 ppm

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

5-weeks

Frequency of treatment:

not applicable

Remarks:

Doses / Concentrations:
100, 500, 2500 ppm
Basis:
nominal in water

No. of animals per sex per dose:

10 males and 10 females rats

Control animals:

yes

Details on study design:

ERKANTOL BXG was diluted once a week in deionized water in the following concentrations: 0 (control), 100, 500 and 2500 ppm by reference to the amount of 22% of test substance.
It was not advisable to use tap water, because with high concentrations it presented a phenomenon of turbidity with precipitation.
The concentrations were chosen based on the results of a pre-test only for a period of one week.
For each dose, the test group consisted of 10 male rats and 10 female rats.
The test substance was diluted once a week in deionized water. To dissolve the deposits present at the bottom, it was heated to approx. 50 ° C. The control subjects received only de-ionized water.
At the beginning of the test, the male and female rats were sorted into two weight groups (light, heavy) and divided into groups corresponding doses based on a random list. Finally, the animals were numbered consecutively.

Positive control:

not applicable

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: 2 times a day on weekdays and once a day on weekends and holidays

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: 2 times a day on weekdays and once a day on weekends and holidays

BODY WEIGHT: Yes
- Time schedule for examinations: at the beginning of the test and weekly

FOOD CONSUMPTION: Yes
The daily food intake was determined by weighing again the amount of uneaten food.

WATER CONSUMPTION: Yes
- Time schedule for examinations: The daily intake of water was determined by measuring the amount of new non-drinking water.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: during the 4th week of test
- Anaesthetic used for blood collection: Yes , ether
- Animals fasted: Yes
- How many animals: 5 males and 5 females of each group
- Erythrocyte count and leukocyte count: counting by Coulter Counter, number of platelets: measurement by TOA PL 100 system, hemoglobin concentration and mean cell volume (MCV): Measurement with a Coulter counter, hematocrit, mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC): Calculation with a Coulter counter, blood count based on smear (modified Wright staining method) thromboplastin time test method according HEENE Hepato-Quick (1974).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: during the 4th week of test
- Animals fasted: Yes
- How many animals: 5 males and 5 females of each group. Due to the suspicious nature of the results, in the fifth week of the test, we measured the concentration of inorganic phosphate on the 5 male rats and the remaining five female rats from each group.
- Enzyme activity in plasma: Alkaline phosphates (ALP), Glutamate-Oxaloacetate-Transaminase (GOT) and glutamate-pyruvate-transaminase (GPT), as recommended by the Deutsche Gesellschaft für Klinische Chemie (1972)
- Substrates in plasma: Creatinine according FABINY and ERTINGSHAUSEN (1971); Urea and BERGMEYER by Gutmann (1974); Glucose by SCHMIDT (1961); Cholesterol in TRINDER (1969); Bilirubin according WAHLEFELD et al. (1972); total albumin according WEICHSELBAUM (1946).
- Electrolytes in serum:Sodium, potassium and calcium determination by flame photometry; Chloride according LANGE LANGE und BORNER (1972);
Inorganic phosphate according DALY and ERTINGSHAUSEN (1972).

URINALYSIS: Yes
- Time schedule for collection of urine: during the 4th week of test. The urine was collected for periods of approx. 16 hours.
- Animals fasted: Yes
- Urinalysis: Semi-quantitative: Glucose, blood, albumin, bilirubin, ketones and pH using sticks Bili-Labstix; Urobilinogènes with Urobilistix, microscopic examinations of sediment after centrifugation of the urine sample; Quantitative: volumes over time; Albumin according RICHTERICH (1968) Specific gravity by digital density meter (DMA 45, Paar Heraeus soc.).

NEUROBEHAVIOURAL EXAMINATION: No data

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Autopsy of rats died during the test:
The rats died during the treatment period were dissected and subjected to macroscopic evaluation. The organs and tissues listed below were treated with Bouin's solution.

Autopsy of rats killed after the test:
All surviving rats were anesthetized with diethyl ether and killed by exsanguination. During the autopsy, the animals were submitted to a pathologico-anatomical examination. The following organs were weighed: brain, heart, testis, lung, liver, spleen, adrenal glands and kidneys.
Preparations of the following organs of all subjects in all groups were treated in a Bouin's solution: aorta, eyes, intestines (duodenum, jejunum, ileum, cecum, colon, rectum), femoral block with skeletal muscle and sciatic nerve, brain, bladder, heart, testis, pituitary gland, salivary gland, lung, lymph nodes (mesenteric and cervical), stomach, spleen, epididymis, adrenal glands, esophagus, ovary, pancreas, prostate, seminal vesicle, the thyroid with parathyroid glands, sternum, thymus, trachea and uterus and all macroscopically visible changes. The liver and kidneys of 5 males and 5 females in each group were also preserved in Bouin's solution. Pieces of liver of these subjects were also preserved by fixation with formalin-calcium. The liver, kidneys and abdominal adipose tissue of other animals were frozen for possible future evaluation.

HISTOPATHOLOGY: Yes
Histopathological examination of the bones (femur, sternum with marrow), heart, liver, spleen, adrenal glands, kidneys and thyroid with parathyroid glands was performed for 5 male rats and five female rats of groups 0 and 2500 ppm.

Other examinations:

no other examination

Statistics:

- Calculation of the arithmetic mean value of each group, the standard deviation , the confidence limits for the confidence level 95% and 99%.
- Comparison of values ​​for the test item-treated and the control groups performed with the significance test (U-test) (MANN, WHITNEY and Wilcoxon (significance thresholds 5% and 1%)).

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Body weight was not affected by the test item in male given 100 and 500 ppm and in females whatever the dose level. Mean body weight gain was slightly delayed in males of group 2 500 ppm (not statistically significant).

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

Food and water intake, general behaviour and mortality were not affected by treatment with the test item. Body weight gain was slightly reduced only in male rats given 2500 ppm (up to 6% v.s. control, not statistically significant).
Up to 2500 ppm, hematological, plasma chemistry and urinary parameters were not affected by the test item.
No test item-related effects were reported at necropsy or during histopathological examinations up to the highest dose level.

Dose descriptor:

NOAEL

Effect level:

2 500 ppm

Based on:

test mat.

Sex:

male/female

Remarks on result:

other: no effects observed

Dose descriptor:

NOAEL

Effect level:

317.05 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Remarks on result:

other: no effects observed

Dose descriptor:

NOAEL

Effect level:

335.22 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Remarks on result:

other: no effects observed

Critical effects observed:

not specified

Table: Results

	DOSE ppm
	0	100	500	2500
MALE FOOD INTAKE g/animal/day	20	20	20	20
FEMALE FOOD INTAKE g/animal/day	15	15	16	16

	DOSE ppm
	0	100	500	2500
MALE WATER CONSUMPTION ml/animal/day	26	24	24	24
MALE TEST COMPOUND INTAKE mg/animal/day mg/kg b.w./day		2.36 11.99	11.87 61.36	59.88 317.05
FEMALE WATER CONSUMPTION ml/animal/day	22	20	18	19
FEMALE TEST COMPOUND INTAKE mg/animal/day mg/kg b.w./day		2.05 14.57	9.15 64.78	46.72 335.22

Haematology 4 weeks
Dose ppm	LEU GIGA/L	ERY TERA/L	HGB G/L	MCV FL	THROM GIGA/L	THP SEC	HCT L/L	MCH PG/E	MCHC G/L
Male 0 100 500 2500	10.2 10.6 9.8 10.3	7.43 7.62 7.52 7.69	154 152 153 155	61 60 62 61	1048 1098 1174 1120	30.5 30.4 31.4 29.3	0.46 0.46 0.47ns 0.47**	20.7 19.9 20.3 20.2	338 330** 328* 330ns
Female 0 100 500 2500	8.1 7.7 7.0 7.3	7.43 7.29 7.45 7.42	151 146 151 151	62 61 61 61	1098 1027 1079 1094	28.7 29.4 30.5 29.2	0.46 0.44 0.46 0.45	20.3 20.0 20.2 20.4	330 329 331 332

 

Differential blood count (%) 4 weeks
Dose ppm	BASO	EOSIN	IPC JGL	STAB	SEGM	LYM	MONO	PLAS
Male 0 100 500 2500	0.0 0.0 0.0 0.0	0.2 0.4 0.4 0.2	0.0 0.0 0.0 0.0	0.0 0.0 0.0 0.0	10.2 8.8 7.6** 7.0*	89.6 90.8 92.0* 92.8ns	0.0 0.0 0.0 0.0	0.0 0.0 0.0 0.0
Female 0 100 500 2500	0.0 0.0 0.0 0.0	0.8 0.4 0.8 0.4	0.0 0.0 0.0 0.0	0.0 0.0 0.0 0.0	6.0 5.8 8.8 8.2	93.2 93.8 90.4 91.4	0.0 0.0 0.0 0.0	0.0 0.0 0.0 0.0

 

Clinical chemistry 4 weeks
Dose ppm	ALP U/L	GOT U/L	GPT U/L	BILI µMOL/L	PROT/P G/L	UREA MMOL/L	CREA µMOL/L	CHOL MMOL/L	GLUC MMOL/L	AN.PH MMOL/L	NA MMOL/L	K MMOL/L	CA MMOL/L	CL MMOL/L
Male 0 100 500 2500	497 437** 430ns 587	63.1 47.8 44.8* 46.5	54.1 50.8 57.3 49.2	2.1 2.1 2.4 2.4	54.7 53.5 54.6 56.3	7.72 7.36 7.72 8.36	40 48 59 43	1.98 2.03 1.91 1.91	5.49 5.51 5.33 5.46	2.48 2.39 2.21** 2.06**	140 139 139 143	5.0 5.2 4.9 4.8	2.75 2.78 2.67 2.76	102 101 103 103
Female 0 100 500 2500	316 321 305 337	43.4 48.5 42.3 52.0	45.7 51.6 52.6 45.5	2.0 2.2 2.2 1.9	57.6 54.6 54.4 53.0	8.40 8.19 7.37* 9.27	54 42 47 45	2.02 1.71* 1.72 1.72	5.22 5.33 5.66 5.18	2.03 1.87 1.71* 1.57**	141 140 140 141	4.4 4.5 4.6 4.2	2.73 2.73 2.68 2.67	102 103 103 104

Quantitative urinalyses 4 weeks
Dose ppm	PROTU MG/16H	VOL ML	SP.GR. G/L
Male 0 100 500 2500	12.35 9.46 7.71 6.48**	35 21 13** 6**	1004 1007 1013 1030**
Female 0 100 500 2500	2.27 1.22ns 1.35* 1.23*	11 5 4* 4*	1010 1024 1034** 1037

Absolute organ weights (mg)
Dose ppm	BODY-W (G)	HEART	LUNG	LIV	SPLE	KIDN	ADRE	TEST	BRAIN
Male 0 100 500 2500	242 243 240 230	783 765 756 711	1032 1040 1075 988	9837 10474 10414 9747	484 482 492 474	1638 1785* 1682 1714	39 40 39 41	2915 2921 2991 2994	1660 1653 1667 1633
Female 0 100 500 2500	173 162 166 162	592 590 588 570	868 806 838 792*	7516 6952 7245 7341	408 379 366 373	1237 1218 1313 1330	58 60 57 60		1617 1652 1658 1652

Relative organ weights (mg/100g)
Dose ppm	BODY-W (G)	HEART	LUNG	LIV	SPLE	KIDN	ADRE	TEST	BRAIN
Male 0 100 500 2500	242 243 240 230	324 315 317 309	427 428 450 430	4063 4298 4328 4232	201 198 207 205	679 734* 700 746*	16 17 17 18	1213 1204 1256 1305	690 683 702 712
Female 0 100 500 2500	173 162 166 162	342 366 357 352	504 500 507 488	4350 4291 4369 4514	236 232 222 229	717 756 796** 819**	33 37 35 37		940 1027* 1008 1023**

Ns : not significant

Significant differences compared to the control group are marked with an asterisk (*) p ≤ 0.05 by two asterisks (**) for p ≤ 0.01.

Conclusions:

Under the test conditions, the test item Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda induced only a slight reduction of body weight gain in males at the dose of 2500 ppm. In the absence of other test item-related changes, this slight variation is considered of no toxicological relevance. The NOAEL was therefore considered to be 2500 ppm (equivalent to 317.05 and 335.22 mg/kg b.w./day in males and females, respectively). Therefore no classification as STOT-RE is required according to the Regulation (EC) 1272/2008 (CLP) and as harmful after repeated dose exposure according to the Directive 67/548/EEC.

Executive summary:

In a repeated dose toxicity study by the oral route, the test item was administered in drinking water to rats (10 animals/sex/dose) at doses of 0; 100; 500 and 2500 ppm during 5 weeks.

Food and water intake, general behaviour and mortality were not affected by treatment. Body weight gain was slightly reduced only in male rats administered 2500 ppm (not statistically significant) but this variation was considered of no toxicological relevance owing to its minimal magnitude (maximum 6% v.s. control) and absence of associated test item-related variations. Up to 2500 ppm, hematological, plasma chemistry and urinary parameters were not affected by the test item. No test item-related effects were reported at necropsy or during histopathological examinations up to the highest dose level.

Based on the results of this study, 2500 ppm of test item, which corresponds to 317.05 and 335.22 mg/kg b.w./day in males and females, respectively, was established as the No Observed Adverse Effect Level (NOAEL). Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

317.05 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Klimisch 2

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is sufficiently documented. OECD Guideline study; analytical purity of test substance: 68,3%.

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K Thomae GmbH
- Age at study initiation: about 7 weeks
- Weight at study initiation: males: mean = 230 g; females: mean = 177 g
- Housing: single
- Diet: ad libitum during post exposure observation period
- Water: ad libitumduring post exposure observation period

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

other: Inhalation: dust aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

air

Remarks on MMAD:

MMAD / GSD: 0.8 mg/m³: 3.3 µm
8 mg/m³: 3.3 µm
80 mg/m³: 3.1 µm

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: aerodynamic exposure apparatus (INA 60, Volume ca. 90 L, BASF AG)
- Method of holding animals in test chamber: The rats were restrained in exposure tubes (glass tubes), their snouts projected into the inhalation chamber and they thus inhaled the dust aerosol.
- System of generating particulates/aerosols: brush generator
- Temperature, humidity, pressure in air chamber: 22°C, 50% humidity
- Method of particle size determination: cascade imapactor

TEST ATMOSPHERE
- Brief description of analytical method used: Spectrophotometric analysis
- Samples taken from breathing zone: yes

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

no data available

Duration of treatment / exposure:

6 hours per day for 28 days

Frequency of treatment:

5 days per week

Remarks:

Doses / Concentrations:
0, 0.8, 8, 80 mg/m³
Basis:
nominal conc.

Remarks:

Doses / Concentrations:
0, 0.86, 8.1, 82 mg/m³
Basis:
analytical conc.

No. of animals per sex per dose:

5
groups of female rats (5 animals) were hold without further exposure for an about 4-week post exposure

Control animals:

yes, concurrent vehicle

Details on study design:

no data available

Positive control:

no data available

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: mortality, behavior and state of health

DETAILED CLINICAL OBSERVATIONS: No

OPHTHALMOSCOPIC EXAMINATION: No

BODY WEIGHT: Yes
- Time schedule for examinations: each week

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 28 day exposure period and after post-exposure observation period
- Anaesthetic used for blood collection: No data
- Animals fasted: No
- How many animals: all
- Parameters checked: leukocytes, erythrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, differential blood count, thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 28 day exposure period and after post-exposure observation period
- Animals fasted: No
- How many animals: all
- Parameters checked: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, sodium, potassium, chloride, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

None

Statistics:

Clinical examinations
Means and standard deviation were calculated for the variables (body weight and body weight gain/body weight change) of the animais of each test group for the statistical evaluation of the study. Statistical relevance was established using methods of ANOVA and DUNNETT.
Clinical chemistry and hematology
Mean and standard deviation were calculated for each test group and tabulated together with the individual values.
Except of the differential blood count, a non-parametric one-way analysis of variance is done via the Kruskal-Wallis-h-test. If the resulting p-value is equal or less than 0.05 a pairwise comparison of each dose group with the control group was carried out. This comparison is done using the Mann-Whitney-U-test for the hypotheses of equal medians.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

statistically significant retarded body weight change in male rats

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

effects observed, treatment-related

Details on results:

CLINICAL SIGNS AND MORTALITY:
80 mg/m³: no substance related effects
8 mg/m³: no substance related effects
0.8 mg/m³: no substance related effects
No mortalities were recorded throughout the exposure period.

BODY WEIGHT AND WEIGHT GAIN:
80 mg/m³: statistically significant retarded body weight change in male rats
8 mg/m³: no substance related effects
0.8 mg/m³: no substance related effects

HAEMATOLOGY:
all groups: no substance related effects

CLINICAL CHEMISTRY:
80 mg/m³: no substance related effects; 8 mg/m³: no substance related effects; 0.8 mg/m³: no substance related effects

ORGAN WEIGHTS:
80 mg/m³: increased absolute and relative lung weights (females and males); in the post exposure observation group a trend towards increased absolute and relative lung weights were still seen
8 mg/m³: increased absolute and relative lung weights which was still apparent in the post exposure observation group
0.8 mg/m³: no substance related effects

GROSS PATHOLOGY:
80 mg/m³: no substance related effects; 8 mg/m³: no substance related effects; 0.8 mg/m³: no substance related effects

HISTOPATHOLOGY:
80 mg/m³: lungs: hypertrophy of goblet cells (male and females), interstitial pneumonitis (increased in grading; males and females), increased connective tissue content (males and females)
nasal cavity (level 1): focal metaplasia of the respiratory epithelium (males and females)
post exposure observation period: increased connective tissue content lungs
8 mg/m³: no substance related effects
0.8 mg/m³: no substance related effects

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

0.86 mg/m³ air (analytical)

Based on:

test mat.

Remarks:

(equivalent to the substance as registered)

Sex:

male/female

Basis for effect level:

other: based on the absence of local effects

Dose descriptor:

LOAEL

Remarks:

local

Effect level:

8.1 mg/m³ air (analytical)

Based on:

test mat.

Remarks:

(equivalent to the substance as registered)

Sex:

male/female

Basis for effect level:

other: based on the effects on lung weight

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

82 mg/m³ air (analytical)

Based on:

test mat.

Remarks:

(equivalent to the substance as registered)

Sex:

male/female

Basis for effect level:

other: based on the absence of toxicologically significant systemic effects

Critical effects observed:

not specified

Conclusions:

In a repeated dose inhalation toxicity study conducted according to OECD Guideline 412 (1990), groups of male and female Wistar rats (5/sex) were exposed by the inhalation route to the test substance as dust aerosol for 6 hours per day on 5 days per week for 4 weeks (28-day test) to head/nose at concentrations of 0, 0.8, 8, 80 mg/m³ (nominal concentrations) or 0, 0.86, 8.1, 82 mg/m³ (analytical concentrations). Based on the results of this study, 82 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) systemic based on the absence of toxicologically significant systemic effects; 0,86 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) local and 8.1 mg/m³ of test item was established as the Lowest Observed Adverse Effect Level (LOAEL) local based on the effects on lung weight. Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Executive summary:

In a repeated dose inhalation toxicity study according to OECD Guideline 412 (1990), groups of male and female Wistar rats (5/sex) were exposed by the inhalation route to the test substance as dust aerosol for 6 hours per day on 5 days per week for 4 weeks (28-day test) to head/nose at concentrations of 0, 0.8, 8, 80 mg/m³ (nominal concentrations) or 0, 0.86, 8.1, 82 mg/m³ (analytical concentrations). Groups of 5 female rats were hold without further exposure for an about 4-week post-exposure observation period to obtain informations on persistency, progression or regression of effects.

No mortality and no clinical signs were reported throughout the study whatever the dose level. Body weight gain was reduced in high dose males only. There were no test item-related effects on hematology and clinical chemistry parameters at any dose levels. No macroscopic findings were reported at necropsy. An increase of absolute and relative lung weights in male animals was observed at 82 mg/m³ and in female animals at 8.1 and 82 mg/m³. Histopathologically, goblet cell hyperplasia was reported in the bronchi of both sexes at 82 mg/m³. Furthermore, the grade of severity of pneumonitis that occurred also in control animals was slightly increased in the animals at 82 mg/m³, as was there a slight increase in the content of connective tissue of the lungs. In the nasal cavity, focal metaplasia of the respiratory epithelium to stratifying epithelium was noted in the animals at 82 mg/m³ at the transition of the stratifying epithelium to respiratory epithelium. After a 4-week recovery period (females only), the lungs of the animals at 82 mg/m³ showed only a slight reparative increase of mature connective tissue. Focal metaplasia in the nasal cavity was evident in only 2 animals at 82 mg/m³.

Based on the results of this study, 82 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) systemic based on the absence of toxicologically significant systemic effects; 0,86 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) local and 8.1 mg/m³ of test item was established as the Lowest Observed Adverse Effect Level (LOAEL) local based on the effects on lung weight.

Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

82 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

Klimisch 2

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is sufficiently documented. OECD Guideline study; analytical purity of test substance: 68,3%.

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K Thomae GmbH
- Age at study initiation: about 7 weeks
- Weight at study initiation: males: mean = 230 g; females: mean = 177 g
- Housing: single
- Diet: ad libitum during post exposure observation period
- Water: ad libitumduring post exposure observation period

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

other: Inhalation: dust aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

air

Remarks on MMAD:

MMAD / GSD: 0.8 mg/m³: 3.3 µm
8 mg/m³: 3.3 µm
80 mg/m³: 3.1 µm

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: aerodynamic exposure apparatus (INA 60, Volume ca. 90 L, BASF AG)
- Method of holding animals in test chamber: The rats were restrained in exposure tubes (glass tubes), their snouts projected into the inhalation chamber and they thus inhaled the dust aerosol.
- System of generating particulates/aerosols: brush generator
- Temperature, humidity, pressure in air chamber: 22°C, 50% humidity
- Method of particle size determination: cascade imapactor

TEST ATMOSPHERE
- Brief description of analytical method used: Spectrophotometric analysis
- Samples taken from breathing zone: yes

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

no data available

Duration of treatment / exposure:

6 hours per day for 28 days

Frequency of treatment:

5 days per week

Remarks:

Doses / Concentrations:
0, 0.8, 8, 80 mg/m³
Basis:
nominal conc.

Remarks:

Doses / Concentrations:
0, 0.86, 8.1, 82 mg/m³
Basis:
analytical conc.

No. of animals per sex per dose:

5
groups of female rats (5 animals) were hold without further exposure for an about 4-week post exposure

Control animals:

yes, concurrent vehicle

Details on study design:

no data available

Positive control:

no data available

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: mortality, behavior and state of health

DETAILED CLINICAL OBSERVATIONS: No

OPHTHALMOSCOPIC EXAMINATION: No

BODY WEIGHT: Yes
- Time schedule for examinations: each week

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 28 day exposure period and after post-exposure observation period
- Anaesthetic used for blood collection: No data
- Animals fasted: No
- How many animals: all
- Parameters checked: leukocytes, erythrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, differential blood count, thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 28 day exposure period and after post-exposure observation period
- Animals fasted: No
- How many animals: all
- Parameters checked: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, sodium, potassium, chloride, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

None

Statistics:

Clinical examinations
Means and standard deviation were calculated for the variables (body weight and body weight gain/body weight change) of the animais of each test group for the statistical evaluation of the study. Statistical relevance was established using methods of ANOVA and DUNNETT.
Clinical chemistry and hematology
Mean and standard deviation were calculated for each test group and tabulated together with the individual values.
Except of the differential blood count, a non-parametric one-way analysis of variance is done via the Kruskal-Wallis-h-test. If the resulting p-value is equal or less than 0.05 a pairwise comparison of each dose group with the control group was carried out. This comparison is done using the Mann-Whitney-U-test for the hypotheses of equal medians.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

statistically significant retarded body weight change in male rats

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

effects observed, treatment-related

Details on results:

CLINICAL SIGNS AND MORTALITY:
80 mg/m³: no substance related effects
8 mg/m³: no substance related effects
0.8 mg/m³: no substance related effects
No mortalities were recorded throughout the exposure period.

BODY WEIGHT AND WEIGHT GAIN:
80 mg/m³: statistically significant retarded body weight change in male rats
8 mg/m³: no substance related effects
0.8 mg/m³: no substance related effects

HAEMATOLOGY:
all groups: no substance related effects

CLINICAL CHEMISTRY:
80 mg/m³: no substance related effects; 8 mg/m³: no substance related effects; 0.8 mg/m³: no substance related effects

ORGAN WEIGHTS:
80 mg/m³: increased absolute and relative lung weights (females and males); in the post exposure observation group a trend towards increased absolute and relative lung weights were still seen
8 mg/m³: increased absolute and relative lung weights which was still apparent in the post exposure observation group
0.8 mg/m³: no substance related effects

GROSS PATHOLOGY:
80 mg/m³: no substance related effects; 8 mg/m³: no substance related effects; 0.8 mg/m³: no substance related effects

HISTOPATHOLOGY:
80 mg/m³: lungs: hypertrophy of goblet cells (male and females), interstitial pneumonitis (increased in grading; males and females), increased connective tissue content (males and females)
nasal cavity (level 1): focal metaplasia of the respiratory epithelium (males and females)
post exposure observation period: increased connective tissue content lungs
8 mg/m³: no substance related effects
0.8 mg/m³: no substance related effects

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

0.86 mg/m³ air (analytical)

Based on:

test mat.

Remarks:

(equivalent to the substance as registered)

Sex:

male/female

Basis for effect level:

other: based on the absence of local effects

Dose descriptor:

LOAEL

Remarks:

local

Effect level:

8.1 mg/m³ air (analytical)

Based on:

test mat.

Remarks:

(equivalent to the substance as registered)

Sex:

male/female

Basis for effect level:

other: based on the effects on lung weight

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

82 mg/m³ air (analytical)

Based on:

test mat.

Remarks:

(equivalent to the substance as registered)

Sex:

male/female

Basis for effect level:

other: based on the absence of toxicologically significant systemic effects

Critical effects observed:

not specified

Conclusions:

In a repeated dose inhalation toxicity study conducted according to OECD Guideline 412 (1990), groups of male and female Wistar rats (5/sex) were exposed by the inhalation route to the test substance as dust aerosol for 6 hours per day on 5 days per week for 4 weeks (28-day test) to head/nose at concentrations of 0, 0.8, 8, 80 mg/m³ (nominal concentrations) or 0, 0.86, 8.1, 82 mg/m³ (analytical concentrations). Based on the results of this study, 82 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) systemic based on the absence of toxicologically significant systemic effects; 0,86 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) local and 8.1 mg/m³ of test item was established as the Lowest Observed Adverse Effect Level (LOAEL) local based on the effects on lung weight. Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Executive summary:

In a repeated dose inhalation toxicity study according to OECD Guideline 412 (1990), groups of male and female Wistar rats (5/sex) were exposed by the inhalation route to the test substance as dust aerosol for 6 hours per day on 5 days per week for 4 weeks (28-day test) to head/nose at concentrations of 0, 0.8, 8, 80 mg/m³ (nominal concentrations) or 0, 0.86, 8.1, 82 mg/m³ (analytical concentrations). Groups of 5 female rats were hold without further exposure for an about 4-week post-exposure observation period to obtain informations on persistency, progression or regression of effects.

No mortality and no clinical signs were reported throughout the study whatever the dose level. Body weight gain was reduced in high dose males only. There were no test item-related effects on hematology and clinical chemistry parameters at any dose levels. No macroscopic findings were reported at necropsy. An increase of absolute and relative lung weights in male animals was observed at 82 mg/m³ and in female animals at 8.1 and 82 mg/m³. Histopathologically, goblet cell hyperplasia was reported in the bronchi of both sexes at 82 mg/m³. Furthermore, the grade of severity of pneumonitis that occurred also in control animals was slightly increased in the animals at 82 mg/m³, as was there a slight increase in the content of connective tissue of the lungs. In the nasal cavity, focal metaplasia of the respiratory epithelium to stratifying epithelium was noted in the animals at 82 mg/m³ at the transition of the stratifying epithelium to respiratory epithelium. After a 4-week recovery period (females only), the lungs of the animals at 82 mg/m³ showed only a slight reparative increase of mature connective tissue. Focal metaplasia in the nasal cavity was evident in only 2 animals at 82 mg/m³.

Based on the results of this study, 82 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) systemic based on the absence of toxicologically significant systemic effects; 0,86 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) local and 8.1 mg/m³ of test item was established as the Lowest Observed Adverse Effect Level (LOAEL) local based on the effects on lung weight.

Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

0.86 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

Klimisch 2

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: oral

One study (Klimish score 2) was available on the substance and was used as a key study:

In a repeated dose toxicity study by the oral route, the test item was administered in drinking water to rats (10 animals/sex/dose) at doses of 0; 100; 500 and 2500 ppm during 5 weeks.

Food and water intake, general behaviour and mortality were not affected by treatment. Body weight gain was slightly reduced only in male rats administered 2500 ppm (not statistically significant) but this variation was considered of no toxicological relevance owing to its minimal magnitude (maximum 6% v.s. control) and absence of associated test item-related variations. Up to 2500 ppm, hematological, plasma chemistry and urinary parameters were not affected by the test item. No test item-related effects were reported at necropsy or during histopathological examinations up to the highest dose level.

Based on the results of this study, 2500 ppm of test item, which corresponds to 317.05 and 335.22 mg/kg b.w./day in males and females, respectively, was established as the No Observed Adverse Effect Level (NOAEL). Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Repeated dose toxicity: inhalation

One study (Klimish score 2) was available on the substance and was used as a key study:

In a repeated dose inhalation toxicity study according to OECD Guideline 412 (1990), groups of male and female Wistar rats (5/sex) were exposed by the inhalation route to the test substance as dust aerosol for 6 hours per day on 5 days per week for 4 weeks (28-day test) to head/nose at concentrations of 0, 0.8, 8, 80 mg/m³ (nominal concentrations) or 0, 0.86, 8.1, 82 mg/m³ (analytical concentrations). Groups of 5 female rats were hold without further exposure for an about 4-week post-exposure observation period to obtain informations on persistency, progression or regression of effects.

No mortality and no clinical signs were reported throughout the study whatever the dose level. Body weight gain was reduced in high dose males only. There were no test item-related effects on hematology and clinical chemistry parameters at any dose levels. No macroscopic findings were reported at necropsy. An increase of absolute and relative lung weights in male animals was observed at 82 mg/m³ and in female animals at 8.1 and 82 mg/m³. Histopathologically, goblet cell hyperplasia was reported in the bronchi of both sexes at 82 mg/m³. Furthermore, the grade of severity of pneumonitis that occurred also in control animals was slightly increased in the animals at 82 mg/m³, as was there a slight increase in the content of connective tissue of the lungs. In the nasal cavity, focal metaplasia of the respiratory epithelium to stratifying epithelium was noted in the animals at 82 mg/m³ at the transition of the stratifying epithelium to respiratory epithelium. After a 4-week recovery period (females only), the lungs of the animals at 82 mg/m³ showed only a slight reparative increase of mature connective tissue. Focal metaplasia in the nasal cavity was evident in only 2 animals at 82 mg/m³.

Based on the results of this study, 82 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) systemicbased on the absence of toxicologically significant systemic effects; 0,86 mg/m³ of test item was established as the No Observed Adverse Effect Level (NOAEL) local and 8.1 mg/m³ of test item was established as the Lowest Observed Adverse Effect Level (LOAEL) local based on theeffects on lung weight.

Therefore, reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the classification criteria of the Regulation (EC) 1272/2008 (CLP) and of the Directive 67/548/EEC.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The selected study is the only study available for the oral route.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
The selected study is the only study available for the inhalation route.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
The selected study is the only study available for the inhalation route.

Justification for classification or non-classification

Harmonized classification:

No harmonized classification is available according to the Regulation (EC) No 1272/2008 including ATP3.

Self classification:

Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified for repeated dose toxicity according to the Regulation (EC) 1272/2008 (CLP) and to the Directive 67/548/EEC.